【作者】 陈蓉；

【导师】 姚裕家；

【作者基本信息】 四川大学 ， 儿科学， 2005， 博士

【摘要】 目的 研究新生大鼠星形胶质细胞（AC）β₁整合素和MMP-2在缺氧/复氧损伤及FDP干预处理后的表达变化和二者之间的相关性,探讨β₁整合素和MMP-2在星形胶质细胞缺氧/复氧损伤中的作用、作用机制和FDP对星形胶质细胞缺氧/复氧损伤β₁整合素和MMP-2表达的影响,为缺氧缺血性脑损伤的机制和治疗提供实验及理论依据。 方法 原代培养成熟纯化的AC随机分成五组: （1）缺氧组:AC在1%氧,无糖无血清的培养基中分别培养12h和24h。 （2）FDP保护组:AC在1%氧,无糖无血清、加入FDP20μl/ml的培养基中分别培养12h和24h。更多还原

【Abstract】 ObjectiveThis paper is to study the expression alterations of β₁ integrin and matrix metalloproteinase-2（MMP-2） in cultured neonatal rat astrocyte （AC） and the correlation between them during experimentalhypoxia-ischemia/reoxygenation（HI/RO） and the intervention of fructose-l,6-disphosphate（FDP） and to explore the effects and mechanism of β₁ integrin and MMP-2 in the astrocytes on the condition of HI/RO and the influence of FDP on the β₁ integrin and MMP-2 expression of astrocytes cell during HI/RO damage, which would provide the laboratory and theory recognition for hypoxia-ischemia brain damage mechanism and management.MethodsThe original cultivating mature purified AC were randomly divided into five groups:（1） the HI group: the astrocytes was exposed to \%O2, without FBS and glucose culture medium for 12h and 24h respectively.（2） FDP protect group: the astrocytes was exposed to 1%O2, without FBS and glucose and added into FDP 20 u I/ml culture medium for 12h and 24h respectively.（3） RO group: the astrocytes subjected to 12h HI, then transferred to ordinary culture for 6h, 12h and 24h respectively.（4） FDP treatment group: the astrocytes subjected to 12h HI, then transferred to ordinary culture and added FDP 20 u I/ml into culture medium 6h, 12h and 24h respectively.（5） control group: normal oxygen cultivated group.The AC body and foot process changes of each sample was observed by light microscope and scanning electron microscope and the expression alterations of β₁ integrin mRNA were determined by real-time PCR and the expression alterations of β₁ integrin and MMP-2 protein were determined by immunofluorescence.Results1. As the time of HI/RO prolonged, AC pattern in morphology changed obviously, mainly showing on the decrease of link between cells, the constracting of cell body, lessening of cell foot process. In FDP protect 12h group and FDP treatment 6h group, AC pattern changed slightly.2. As the time of HI/RO prolonged, the expression level of β₁ integrinmRNA and protein in AC significantly decreased （P<0.05）. The (3j integrin mRNA and protein expression of FDP protect group obviously increased compared with the same period of HI group （P<0.05） and them of FDP treatment group obviously increased compared with the same period of RO group （P<0.05）.3. As the time of HI/RO prolonged, the expression of MMP-2 protein in AC obviously increased in FDP protect group evidently increased compared with the same period of HI group （P<0.05） and their expression in FDP protect group significantly decreased compared with the same period of HI Group （P<0.05） and their expression in FDP treatment group obviously decreased compared with the same period of RO group （P<0.05）.4. The relationship of Pi integrin and MMP-2 protein expression showed a negative correlation during AC HI/RO damage.Conclusion1. During AC HI/RO damage, the expression of Pi integrin in AC decreased and MMP-2 increased and the links between AC were broken, which may affect the normal pattern and function of AC.2. FDP could maintain links between AC and normal pattern of AC and reduce the AC damage from HI/RO through increasing the expression of Pi integrin and decreasing the expression of MMP-2.更多还原